Table 1 Comparative advantages of B. subtilis spore surface display and B. subtilis cell surface display systems versus other display technologies for biotechnological applications

Protein folding efficiency

Circumvents folding issues as proteins are displayed outside the membrane

Proteins are directly expressed on the cell surface, maintaining near-native conformation

Soluble protein systems may require chaperones for correct folding

[22, 25]

Stability under harsh conditions

Highly resistant to heat, desiccation, radiation, UV, and oxidation

Moderate stability; proteins may degrade under extreme conditions

Purified proteins require stabilizers for stability

[26, 27]

Immobilization process

Proteins are naturally immobilized during sporulation

Proteins are anchored on live cells during expression, allowing dynamic interactions

Other systems (e.g., phage, yeast) require genetic engineering and optimization

[22, 28]

High-throughput screening

Suitable for screening in extreme environments due to robustness

Enables real-time functional assays for enzyme engineering

Cell-free and phage systems lack cellular context but offer flexibility

[29,30,31]

Economic feasibility

Cost-effective; large-scale production without special culture conditions

Cost-efficient, but requires fresh cultures for best performance

Purified protein production is costly due to purification steps

[11, 32]

Safety considerations

GRAS (Generally Recognized as Safe) status; suitable for long-term storage

Safe for live-cell vaccines and probiotics

Recombinant systems (e.g., E. coli) may need additional precautions

[33]

Ease of handling

Durable and storable for long periods without refrigeration

Requires continuous culture maintenance to preserve functionality

Purified proteins need cold storage and stabilizers

[11, 34]

Protein functionality

Immobilized proteins retain activity and stability in harsh conditions

Active proteins in a near-native environment for functional assays

Soluble proteins may have high activity but lack durability

[35, 36]

Dynamic vs. static applications

Suitable for static applications like vaccines or long-term catalysts

Enables dynamic applications like continuous substrate processing

Phage and yeast display offer high diversity for functional screening

[11, 37]